OBJECTIVE To explore the effect of respiratory syncytial virus(RSV)infection on the expression of key factors in the epithelial barrier of the human nasal epithelial cells(hNECs)derived from patients with chronic rhinosinusitis with nasal polyps(CRSwNP)and normal control mucosa.METHODS RSV with different multiplicity of infection(MOI)(0.1 and 0.3)infected hNECs derived from patients with CRSwNP(n=21)and normal control mucosa(n=9)for 24 h and 48 h,respectively.To detect the gene expression ZO-1,ZO-2,Claudin-1,Claudin-4,Occludin,E-cadherin and N-cadherin,total RNA was extracted and reverse transcribed into cDNA for real-time fluorescence quantification PCR.RESULTS The relative expression level of ZO-1,ZO-2,Claudin-1,Claudin-4,Occludin,E-cadherin and N-cadherin were decreased in hNECs post RSV infection.However,there was a statistical difference only in hNECs derived from CRSwNP(P<0.05).There was no significant difference in hNECs infected with RSV between eosinophilic CRSwNP(ECRSwNP)and non-eosinophilic CRSwNP(nonECRSwNP).CONCLUSION RSV infection could disrupt the epithelial barrier of the nasal mucosa,and patients with CRSwNP are more severely affected by RSV infection compared to healthy controls.The impact of RSV infection on mucosa between ECRSwNP group and nonECRSwNP group was no significant difference.

Objective To study the mechanical properties of titanium mesh and three-dimensional(3D)-printed metal vertebral body substitutes(VBS)to provide guidance for the selection and structural optimization of artificial vertebral implants in clinical practice.Methods The equivalent elastic modulus,equivalent yield strength,and structural failure mode of titanium mesh and 3D-printed porous,truss,and topologically optimized VBS were systematically investigated using compression tests.Results The elastic modulus of the titanium mesh(2 908.73±287.39 MPa)was only lower than that of the topologically optimized VBS.However,their structural strengths and stabilities were inadequate.The yield strength of the titanium mesh(46.61±4.85 MPa)was only higher than that of the porous VBS and it was the first to yield during compression.The porous VBS was insufficient for use as the vertebral implant owing to its poor mechanical strength(18.14±0.17 MPa-25.79±0.40 MPa).The truss VBS had good elastic modulus(2 477.86±55.19 MPa-2 620.08±194.36 MPa)and strength(77.61±0.50 MPa-88.42±1.07 MPa).However,the structural stability of the truss VBS was insufficient,and instability occurred easily during compression.The topologically optimized VBS had the highest elastic modulus(3 746.28±183.80 MPa)and yield strength(177.43±3.82 MPa)among all the tested VBS types,which could provide improved security and stability for artificial vertebral implant in vivo services.Conclusions Topology optimization results in a high strength and high stability VBS design.Moreover,it provides a large design space and great safety margin to provide increased possibilities for lightweight and new material design of future artificial vertebral implants.

OBJECTIVE: To explore the genetic basis for a Chinese pedigree affected with Branchio-Oto syndrome (BOS). METHODS: A pedigree with BOS which had presented at the Genetics and Prenatal Diagnosis Center of the First Affiliated Hospital of Zhengzhou University in May 2021 was selected as the study subject. Clinical data of the pedigree was collected. Peripheral blood samples of the proband and her parents were collected. Whole exome sequencing (WES) was carried out for the proband. Multiplex ligation-dependent probe amplification (MLPA) was used to verify the result of WES, short tandem repeat (STR) analysis was used to verify the relationship between the proband and her parents, and the pathogenicity of the candidate variant was analyzed. RESULTS: The proband, a 6-year-old girl, had manifested severe congenital deafness, along with inner ear malformation and bilateral branchial fistulae. WES revealed that she has harbored a heterozygous deletion of 2 466 kb at chromosome 8q13.3, which encompassed the EYA1 gene. MLPA confirmed that all of the 18 exons of the EYA1 gene were lost, and neither of her parents has carried the same deletion variant. STR analysis supported that both of her parents are biological parents. Based on the guidelines from the American College of Medical Genetics and Genomics, the deletion was classified as pathogenic (PVS1+PS2+PM2_Supporting+PP4). CONCLUSION The heterozygous deletion of EYA1 gene probably underlay the pathogenicity of BOS in the proband, which has provided a basis for the clinical diagnosis.
Humans ; Female ; Pregnancy ; Child ; Pedigree ; Family ; Parents ; Chromosomes, Human, Pair 3 ; Exons ; Nuclear Proteins/genetics* ; Protein Tyrosine Phosphatases ; Intracellular Signaling Peptides and Proteins/genetics*

OBJECTIVE: To explore the genetic basis for a Chinese pedigree featuring congenital profound syndromic deafness and chronic constipation, and provide prenatal diagnosis for a high-risk fetus. METHODS: Whole-exome sequencing was carried out to analyze the sequences of genes associated with hereditary deafness, and multiplex ligation-dependent probe amplification (MLPA) was used to verify the candidate variant in the proband's parents and the fetus. RESULTS: The proband was found to have harbored a heterozygous deletion of SOX10, a pathogenic gene associated with Waardenburg syndrome type 4C (WS4C). The same deletion was found in her mother (with profound syndromic deafness and chronic constipation) and the fetus, but not in her father with normal hearing. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG) and Association for Molecular Pathology (AMP), the SOX10 gene deletion was predicted to be a pathogenic variant (PVS1+PM2_Supporting+PP1+PP4). CONCLUSION The pedigree was diagnosed with WS4C, which has conformed to an autosomal dominant inheritance. Deletion of the entire SOX10 gene, as a loss-of-function variant, probably underlay its pathogenesis. Above finding has facilitated genetic counseling and prenatal diagnosis for this family.
Humans ; Female ; Pregnancy ; Pedigree ; Waardenburg Syndrome/genetics* ; East Asian People ; Genetic Testing ; Prenatal Diagnosis ; Hearing Loss, Sensorineural/genetics* ; Deafness/genetics* ; Mothers ; Constipation/genetics* ; Mutation ; SOXE Transcription Factors/genetics*

OBJECTIVE To investigate the inhibitory effects of ursolic acid on interleukin-6 （IL-6）-mediated invasion and migration of breast cancer MDA-MB-231 cells （hereinafter referred to as “231 cells”）. METHODS The effects of 20， 40， 80， 160 and 320 µmol/L ursolic acid on the proliferation rate of 231 cells were measured by CCK-8 method. The breast cancer 231 cells were divided into control group， model group and administration group. The migration and invasion abilities of cells were detected by scratch assay and Transwell assay. Real-time quantitative polymerase chain reaction （q-PCR） assay and Western blot assay were used to detect the mRNA and protein expressions of epithelial-mesenchymal transition-related makers such as E cadherin （E-cad）， matrix metalloprotein 2 （MMP2）， MMP9， vimentin （Vim）， CD44 molecule （CD44） and aldehyde dehydrogenase 1 family member A1 （ALDH1A1）. The phosphorylation levels of JAK2 and STAT3 in the Janus kinase 2/signal transducer and activator of transcription 3 （JAK2/STAT3） signaling pathway （in terms of p-JAK2/JAK2 ratio and p-STAT3/STAT3 ratio） were detected by Western blot assay. RESULTS A low concentration of ursolic acid of 20 µmol/L （no significant inhibitory effect on cell proliferation ability） was selected as the subsequent administration concentration. Compared with the control group， the migration and invasion abilities of cells in the model group were significantly enhanced （P＜0.05）； compared with the model group， the migration and invasion abilities of cells in the administered group were significantly reduced （P＜0.05）. Compared with the control group， the relative mRNA and protein expressions of epithelial-mesenchymal transition-related markers MMP9， MMP2， Vim， ALDH1A1 and CD44 were all elevated to different extents， and the mRNA and protein expressions of E-cad were all decreased to different extents in the model group cells， and part of the differences had statistical significance （P＜0.05）， the p-JAK2/JAK2 ratio and p-STAT3/STAT3 ratio were significantly increased in the model group （P＜0.05）； compared with the model group， the expressions of the above indicators were reversed to some extent in the administration group. CONCLUSIONS Ursolic acid blocks the activation of JAK2/STAT3 signaling pathwby the inflammatory factor IL-6， which ultimately interrupts the invasion and metastasis of breast cancer cells.

Humans ; COVID-19 ; Consensus ; SARS-CoV-2 ; China

OBJECTIVE: To explore the genetic basis for a Chinese pedigree affected with congenital deafness pedigree in conjunct with enlarged vestibular aqueduct. METHODS: Whole-exome sequencing was carried out for the proband to analyze the genes associated with hereditary deafness. Candidate variant was verified by Sanger sequencing of the proband's parents and her younger brother. RESULTS: The proband was found to harbor compound heterozygous variants including c.748dupG (p.Asp250Glyfs*30Asn) (pathogenic, PVS1+PM2+PP4) and c.879C>A (p.Ser293Arg) (likely pathogenic, PM2+PM3+PP1+PP4) of the FOXI1 gene, which has been associated with enlarged vestibular aqueduct (OMIM 600791). Both variants were unreported previously. The variants were respectively inherited from proband's parents whom had normal hearing. Her younger brother was heterozygous for the c.748dupG variant but also had normal hearing. CONCLUSION The compound heterozygous variants of the FOXI1 gene probably underlay the pathogenicity of congenital deafness and enlarged vestibular aqueduct in the proband. The co-segregation of the two variants with the hearing loss has facilitated genetic counseling and prenatal diagnosis for this pedigree.
China ; Deafness/genetics* ; Female ; Forkhead Transcription Factors/genetics* ; Hearing Loss, Sensorineural ; Humans ; Male ; Mutation ; Pedigree ; Pregnancy ; Vestibular Aqueduct/abnormalities*

Objective:To explore the genetic pathogenicity for a Chinese pedigree affected with severe syndromic deafness.Methods:High-throughput sequencing was carried out to analyze the 415 genes associated with hereditary deafness in the proband who has hearing loss in association with optic atrophy and hyperglycemia. Candidate variants were verified by Sanger sequencing of the proband, her parents and the fetus.Results:The proband was found to harbor compound heterozygous variants of WFS1 gene, namely c. 2389G>A (p.Asp797Asn) and c. 2345C>T (p.Pro782Leu), which was known to underlie Wolfram syndrome 1. The proband′s parents had normal hearing and were both heterozygous carriers for the above variants. The fetus was found to harbor the same compound heterozygous variants and was predicted to have a high risk. Conclusion:The compound heterozygous variants of c. 2389G>A and c. 2345C>T of the WFS1 gene probably underlay the pathogenesis of hearing loss in the proband. Above finding has facilitated genetic counseling and prenatal diagnosis for this family.

OBJECTIVE: To explore the genetic basis of a Chinese pedigree affected with progressive non-syndromic sensorineural hearing loss. METHODS: High-throughput DNA sequencing was carried out to analyze 415 genes associated with hereditary deafness in the proband. Sanger sequencing was carried out to verify the suspected variants among her family members. RESULTS: The proband was found to carry a heterozygous c.842T>A (p.Ile281Asn) variant of the POU4F3 gene. The same variant was found among all other patients from the pedigree including the proband's mother, brother, aunt and maternal grandfather, but not among those with normal hearing. Based on the standards and guidelines of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology, the c.842T>A(p.Ile281Asn) variant of the POU4F3 gene was predicted as likely pathogenic (PM2+PM5+PP1+PP3+PP4). CONCLUSION A Chinese pedigree affected by a rare type autosomal dominant deafness-15 (DFNA15) due to a novel c.842T>A (p.Ile281Asn) variant of the POU4F3 gene was identified. The result has facilitated genetic counseling and risk assessment for the pedigree.
China ; Deafness/genetics* ; Female ; Genetic Testing ; Hearing Loss, Sensorineural/genetics* ; Humans ; Male ; Mutation ; Pedigree

OBJECTIVE: To explore the genetic basis for a pedigree affected with congenital sensorineural deafness. METHODS: High-throughput sequencing was carried out to analyze the coding regions of 415 genes associated with hereditary deafness in the proband. Suspected variants were verified by PCR amplification and Sanger sequencing of her parents and sister. RESULTS: The proband was found to have carried a heterozygous c.5131G>A (p.Val1711Ile) variant of the CDH23 gene and a heterozygous c.2884C>T(p.Arg962Cys) variant of the PCDH15 gene, which were respectively inherited from her mother and father. Her sister (with normal hearing) was also heterozygous for the c.5131G>A (p.Val1711Ile) variant of the CDH23 gene but not the c.2884C>T (p.Arg962Cys) variant of the PCDH15 gene. Based on the guidelines of the American College of Medical Genetics and Genomics, both variants were predicted to be likely pathogenic (PS1+PM2+PP3+PP4). CONCLUSION The c.5131G>A (p.Val1711Ile) variant of the CDH23 gene and c.2884C>T (p.Arg962Cys) variant of the PCDH15 gene probably underlay the pathogenesis of Usher syndrome type 1D/F in this pedigree.
Female ; Heterozygote ; High-Throughput Nucleotide Sequencing ; Humans ; Mutation ; Pedigree ; Usher Syndromes/genetics*